Abstract:
:Transcription activator-like effector (TALE) nuclease (TALEN) is a site-specific nuclease, which can be freely designed and easily constructed. Numerous methods of constructing TALENs harboring different TALE scaffolds and repeat variants have recently been reported. However, the functionalities of structurally different TALENs have not yet been compared. Here, we report on the functional differences among several types of TALENs targeting the same loci. Using HEK293T cell-based single-strand annealing and Cel-I nuclease assays, we found that TALENs with periodically-patterned repeat variants harboring non-repeat-variable di-residue (non-RVD) variations (Platinum TALENs) showed higher activities than TALENs without non-RVD variations. Furthermore, the efficiencies of gene disruption mediated by Platinum TALENs in frogs and rats were significantly higher than in previous reports. This study therefore demonstrated an efficient system for the construction of these highly active Platinum TALENs (Platinum Gate system), which could establish a new standard in TALEN engineering.
journal_name
Sci Repjournal_title
Scientific reportsauthors
Sakuma T,Ochiai H,Kaneko T,Mashimo T,Tokumasu D,Sakane Y,Suzuki K,Miyamoto T,Sakamoto N,Matsuura S,Yamamoto Tdoi
10.1038/srep03379subject
Has Abstractpub_date
2013-11-29 00:00:00pages
3379issn
2045-2322pii
srep03379journal_volume
3pub_type
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